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Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

Editor-in-Chief: Plebani, Mario

Ed. by Gillery, Philippe / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Payne, Deborah A. / Schlattmann, Peter / Tate, Jillian R.

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1437-4331
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Volume 56, Issue 1

Issues

Determination of reference intervals for urinary steroid profiling using a newly validated GC-MS/MS method

Wilhelmina H.A. de Jong
  • Corresponding author
  • Department of Laboratory Medicine, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands
  • Email
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Edward Buitenwerf
  • Department of Endocrinology, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Alle T. Pranger
  • Department of Laboratory Medicine, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Ineke J. RiphagenORCID iD: http://orcid.org/0000-0001-5500-0145 / Bruce H.R. Wolffenbuttel
  • Department of Endocrinology, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Michiel N. Kerstens
  • Department of Endocrinology, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands
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  • De Gruyter OnlineGoogle Scholar
/ Ido P. Kema
  • Department of Laboratory Medicine, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
Published Online: 2017-06-09 | DOI: https://doi.org/10.1515/cclm-2016-1072

Abstract

Background:

Urinary steroid profiling (USP) is a powerful diagnostic tool to asses disorders of steroidogenesis. Pre-analytical factors such as age, sex and use of oral contraceptive pills (OCP) may affect steroid hormone synthesis and metabolism. In general, USP reference intervals are not adjusted for these variables. In this study we aimed to establish such reference intervals using a newly-developed and validated gas chromatography with tandem mass spectrometry detection method (GC-MS/MS).

Methods:

Two hundred and forty healthy subjects aged 20–79 years, stratified into six consecutive decade groups each containing 20 males and 20 females, were included. None of the subjects used medications. In addition, 40 women aged 20–39 years using OCP were selected. A GC-MS/MS assay, using hydrolysis, solid phase extraction and double derivatization, was extensively validated and applied for determining USP reference intervals.

Results:

Androgen metabolite excretion declined with age in both men and women. Cortisol metabolite excretion remained constant during life in both sexes but increased in women 70–79 years of age. Progesterone metabolite excretion peaked in 30–39-year-old women and declined afterwards. Women using OCP had lower excretions of androgen metabolites, progesterone metabolites and cortisol metabolites. Method validation results met prerequisites and revealed the robustness of the GC-MS/MS method.

Conclusions:

We developed a new GC-MS/MS method for USP which is applicable for high throughput analysis. Widely applicable age and sex specific reference intervals for 33 metabolites and their diagnostic ratios have been defined. In addition to age and gender, USP reference intervals should be adjusted for OCP use.

This article offers supplementary material which is provided at the end of the article.

Keywords: GC-MS/MS; oral contraceptive pills; reference intervals; urinary steroid profile

References

  • 1.

    Miller WL, Auchus RJ. The molecular biology, biochemistry, and physiology of human steroidogenesis and its disorders. Endocr Rev 2011;32:81–151.PubMedWeb of ScienceCrossrefGoogle Scholar

  • 2.

    Miller WL, Bose HS. Early steps in steroidogenesis: intracellular cholesterol trafficking. J Lipid Res 2011;52:2111–35.PubMedWeb of ScienceCrossrefGoogle Scholar

  • 3.

    Larsen P, Kronenberg H, Melmed S, Polonsky K. Effects of glucocorticoids. In: Williams textbook of endocrinology, 10th ed. Philadelphia: W.B Saunders Company, 2003:503–6.Google Scholar

  • 4.

    Krone N, Hughes BA, Lavery GG, Stewart PM, Arlt W, Shackleton CH. Gas chromatography/mass spectrometry (GC/MS) remains a pre-eminent discovery tool in clinical steroid investigations even in the era of fast liquid chromatography tandem mass spectrometry (LC/MS/MS). J Steroid Biochem Mol Biol 2010;121: 496–504.Web of SciencePubMedCrossrefGoogle Scholar

  • 5.

    Wudy SA, Hartmann MF. Gas chromatography-mass spectrometry profiling of steroids in times of molecular biology. Horm Metab Res 2004;36:415–22.CrossrefPubMedGoogle Scholar

  • 6.

    Taylor NF. Urinary steroid profiling. Methods Mol Biol 2013;1065:259–76.Web of SciencePubMedCrossrefGoogle Scholar

  • 7.

    Wolthers BG, Kraan GP. Clinical applications of gas chromatography and gas chromatography-mass spectrometry of steroids. J Chromatogr A 1999;843:247–74.CrossrefPubMedGoogle Scholar

  • 8.

    Kerstens MN, Guillaume CP, Wolthers BG, Dullaart RP. Gas chromatographic-mass spectrometric analysis of urinary glycyrrhetinic acid: an aid in diagnosing liquorice abuse. J Intern Med 1999;246:539–47.CrossrefPubMedGoogle Scholar

  • 9.

    Berruti A, Baudin E, Gelderblom H, Haak HR, Porpiglia F, Fassnacht M, et al. Adrenal cancer: ESMO Clinical Practice Guidelines for diagnosis, treatment and follow-up. Ann Oncol 2012;23(Suppl 7):vii, 131–8.Web of ScienceGoogle Scholar

  • 10.

    Kerkhofs TM, Kerstens MN, Kema IP, Willems TP, Haak HR. Diagnostic value of urinary steroid profiling in the evaluation of adrenal tumors. Horm Cancer 2015;6:168–75.PubMedWeb of ScienceCrossrefGoogle Scholar

  • 11.

    Arlt W, Biehl M, Taylor AE, Hahner S, Libé R, Hughes BA, et al. Urine steroid metabolomics as a biomarker tool for detecting malignancy in adrenal tumors. J Clin Endocrinol Metab 2011;96:3775–84.Web of ScienceCrossrefPubMedGoogle Scholar

  • 12.

    Grondal S, Eriksson B, Hagenas L, Werner S, Curstedt T. Steroid profile in urine: a useful tool in the diagnosis and follow up of adrenocortical carcinoma. Acta Endocrinol (Copenh) 1990;122:656–63.PubMedGoogle Scholar

  • 13.

    Van Renterghem P, Van Eenoo P, Geyer H, Schanzer W, Delbeke FT. Reference ranges for urinary concentrations and ratios of endogenous steroids, which can be used as markers for steroid misuse, in a Caucasian population of athletes. Steroids 2010;75:154–63.CrossrefWeb of ScienceGoogle Scholar

  • 14.

    Weykamp CW, Penders TJ, Schmidt NA, Borburgh AJ, van de Calseyde JF, Wolthers BJ. Steroid profile for urine: reference values. Clin Chem 1989;35:2281–4.PubMedGoogle Scholar

  • 15.

    Martinez-Brito D, Correa Vidal MT, de la Torre X, Garcia-Mir V, Ledea Lozano O, Granda Fraga M. Reference ranges for the urinary steroid profile in a Latin-American population. Drug Test Anal 2013;5:619–26.Web of ScienceCrossrefGoogle Scholar

  • 16.

    Chan AO, Taylor NF, Tiu SC, Shek CC. Reference intervals of urinary steroid metabolites using gas chromatography-mass spectrometry in Chinese adults. Steroids 2008;73:828–37.PubMedWeb of ScienceCrossrefGoogle Scholar

  • 17.

    Shackleton CH. Mass spectrometry in the diagnosis of steroid-related disorders and in hypertension research. J Steroid Biochem Mol Biol 1993;45:127–40.CrossrefGoogle Scholar

  • 18.

    Scholtens S, Smidt N, Swertz MA, Bakker SJ, Dotinga A, Vonk JM, et al. Cohort profile: LifeLines, a three-generation cohort study and biobank. Int J Epidemiol 2015;44:1172–80.CrossrefPubMedWeb of ScienceGoogle Scholar

  • 19.

    Wielders JP, Roelofsen-de Beer RJ, Boer AK, de Jong WH, Mohrmann K, Mulder AH, et al. Validatie en verificatie van onderzoeksprocedures in medische laboratoria. Ned Tijdschr Klin Chem 2017;42:25–36.Google Scholar

  • 20.

    Kirschbaum C, Kudielka BM, Gaab J, Schommer NC, Hellhammer DH. Impact of gender, menstrual cycle phase, and oral contraceptives on the activity of the hypothalamus-pituitary-adrenal axis. Psychosom Med 1999;61:154–62.PubMedCrossrefGoogle Scholar

  • 21.

    Bulbrook RD, Herian M, Tong D, Hayward JL, Swain MC, Wang DY. Effect of steroidal contraceptives on levels of plasma androgen sulphates and cortisol. Lancet 1973;1:628–31.PubMedGoogle Scholar

  • 22.

    Labrie F, Belanger A, Cusan L, Gomez JL, Candas B. Marked decline in serum concentrations of adrenal C19 sex steroid precursors and conjugated androgen metabolites during aging. J Clin Endocrinol Metab 1997;82:2396–402.CrossrefPubMedGoogle Scholar

  • 23.

    Parker CR Jr, Slayden SM, Azziz R, Crabbe SL, Hines GA, Boots LR, et al. Effects of aging on adrenal function in the human: responsiveness and sensitivity of adrenal androgens and cortisol to adrenocorticotropin in premenopausal and postmenopausal women. J Clin Endocrinol Metab 2000;85:48–54.PubMedGoogle Scholar

  • 24.

    Schäfer HH, de Villiers JD, Sivukhina E, Lewis J, Wande D, Perembe B, et al. Altered homeostasis of systemic glucocorticoids as related to obesity, glucose tolerance, and smoking. Horm Metab Res 2013;45:245–51.PubMedWeb of ScienceGoogle Scholar

  • 25.

    Guo X, Lankmayr E. Hyphenated techniques in gas chromatography. In: Mustafa Ali Mohd, editor. Advanced gas chromatography — progress in agricultural, biomedical and industrial applications. InTech, 2012. ISBN: 978-953-51-0298-4. Available from: http://www.intechopen.com/books/advanced-gaschromatography-progress-in-agricultural-biomedical-and-industrial-applications/hyphenated-techniques-ingas-chromatography.

  • 26.

    Marcos J, Renau N, Casals G, Segura J, Ventura R, Pozo OJ. Investigation of endogenous corticosteroids profiles in human urine based on liquid chromatography tandem mass spectrometry. Anal Chim Acta 2014;812:92–104.PubMedCrossrefWeb of ScienceGoogle Scholar

About the article

Corresponding author: Wilhelmina H.A. de Jong, PhD, Department of Laboratory Medicine, University of Groningen, University Medical Center Groningen, Hanzeplein 1, EA61, PO-Box 30.001, 9700 RB Groningen, The Netherlands, Phone: +0031-50-3617015, Fax: +050-3611746


Received: 2016-11-24

Accepted: 2017-04-20

Published Online: 2017-06-09

Published in Print: 2017-11-27


Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

Research funding: None declared.

Employment or leadership: None declared.

Honorarium: None declared.

Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.


Citation Information: Clinical Chemistry and Laboratory Medicine (CCLM), Volume 56, Issue 1, Pages 103–112, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/cclm-2016-1072.

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